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Literature summary for 3.2.1.133 extracted from
- Modulation of hydrolysis and transglycosylation activity of Thermus maltogenic amylase by combinatorial saturation mutagenesis (2008), J. Microbiol. Biotechnol., 18, 1401-1407.
Application
| Application | Comment | Organism |
|---|---|---|
| biotechnology | producing enzymes with modified substrate specificity, hydrolysis, and transglycosilation activities, as a way to produce specific functional carbohydrate materials | Thermus sp. |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| cloning and expression in Escherichia coli MC 1061 with p6xHTMX, transformants grown in Luria-Bertani medium with ampicillin at 37°C | Thermus sp. |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A330G/N331C/E332C | F-18, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| A330G/N331G/E332C | C-20, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| A330G/N331G/E332G | G-91, strong reduction of all substrate hydrolyzing activities, higher relative specificity to beta-cyclodextrin than to starch compared to wild-type, lower relative specificity to maltotriose than to acarbose compared to wild-type, transglycosylation: high amount of branched tetraose and pentaose | Thermus sp. |
| A330G/N331G/E332S | G-22, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| A330G/N331P/E332G | C-43, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| A330G/N331V/E332G | G-90, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| A330M/N331G/E332C | B-96, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| A330S/N331A | F-80, strong reduction of all substrate hydrolyzing activities, higher relative specificity to beta-cyclodextrin and pullulan than to starch compared to wild-type, lower relative specificity to maltotriose than to acarbose compared to wild-type, transglycosylation: high amount of branched tetraose and pentaose | Thermus sp. |
| A330S/N331G/E332T | K-37, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodexxtrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| additional information | Random substitutions of amino acid residues Val329, Ala330, Asn331, and Glu332 that are forming an extra sugar-binding space with combinatorial saturation mutagenesis technique: substrate specificity, hydrolysis pattern, and transglycosylation activity of ThMA are modulated by these mutations. Activity assays with substrates: soluble starch, pullulan, beta-cyclodextrin, maltotriose, and acarbose at 60°C, pH 6.0 sodium-acetate buffer | Thermus sp. |
| N331S/E332G | I-69, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| V329A/A330C/N331G/E332V | A-39, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| V329A/A330G/N331V/E332A | G-13, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| V329A/N331L | B-4, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch compared to wild-type | Thermus sp. |
| V329C/N331H/E332R | D-3, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| V329F/A330T/N331G/E332W | I-70, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| V329G/A330L/N331V/E332Y | H-16, slightly higher activity with cyclodextrin, pullulan, and starch, reduced activities with maltotriose, and acarbose, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type, similar transglysocylation pattern as wild-type | Thermus sp. |
| V329I/A330G/N331W | C-56, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| V329S/A330C/N331S/E332P | A-18, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| V329S/A330G/N331D | K-33, strong reduction of all substrate hydrolyzing activities, products of acarbose hydrolization are glucose, maltose, and acarviosine instead of only glucose and carviosine-glucose as in the wild-type reaction, higher relative specificity to beta-cyclodextrin than to starch compared to wild-type, lower relative specificity to maltotriose than to acarbose compared to wild-type, transglycosylation: very little amount of transfer products, with acarbose significant amount of acarviosine and maltose | Thermus sp. |
| V329S/A330G/N331G/E332V | E-74, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
| V329T/A330C/N331T/E332V | E-50, strong reduction of all substrate hydrolyzing activities, lower relative specificity to beta-cyclodextrin and pullulan than to starch and to maltotriose than to acarbose compared to wild-type | Thermus sp. |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| acarbose + H2O | Thermus sp. | - |
glucose + acarviosine-glucose | - |
? |  |
| alpha-(1,4)-glycosidic linked cyclodextrins + H2O | Thermus sp. | main depolymerization of outer amylopectin branches | maltooligosaccharide | - |
? | |
| beta-cyclodextrin + H2O | Thermus sp. | - |
maltooligosaccharide | - |
? | |
| maltotriose + H2O | Thermus sp. | - |
isomaltose + isopanose + panose + branched glucooligosaccharides | transfer products of transglycosylation | ? | |
| pullulan + H2O | Thermus sp. | - |
panose + ? | - |
? | |
| starch + H2O | Thermus sp. | - |
maltooligosaccharide | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Thermus sp. | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| purification of ThMA variants with N-terminal six-histidines by nickel-nitrilotriacetic acid column chromatography | Thermus sp. |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| acarbose + H2O | - |
Thermus sp. | glucose + acarviosine-glucose | - |
? | |
| alpha-(1,4)-glycosidic linked cyclodextrins + H2O | main depolymerization of outer amylopectin branches | Thermus sp. | maltooligosaccharide | - |
? | |
| beta-cyclodextrin + H2O | - |
Thermus sp. | maltooligosaccharide | - |
? | |
| maltotriose + H2O | - |
Thermus sp. | isomaltose + isopanose + panose + branched glucooligosaccharides | transfer products of transglycosylation | ? | |
| pullulan + H2O | - |
Thermus sp. | panose + ? | - |
? | |
| starch + H2O | - |
Thermus sp. | maltooligosaccharide | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| maltogenic alpha-amylase | - |
Thermus sp. |
| Thermus maltogenic amylase | hydrolyzes cyclodextrin faster than starch | Thermus sp. |
| ThMA | - |
Thermus sp. |
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